Department of Pediatric Medicine I, The First Affiliated Hospital of Jiamusi University, No. 348, Dexiang Street, Xiangyang District, Jiamusi City, 154002, Heilongjiang Province, China.
Department of General Surgery I, The First Affiliated Hospital of Jiamusi University, No. 348, Dexiang Street, Xiangyang District, Jiamusi City, 154002, Heilongjiang Province, China.
Mol Cell Biochem. 2021 Feb;476(2):1063-1074. doi: 10.1007/s11010-020-03972-8. Epub 2020 Nov 10.
Previous studies have reported the important roles of long non-coding RNAs (lncRNAs) in acute respiratory distress syndrome (ARDS). Here, we focus on the role and regulatory mechanism of lncRNA SNHG5 in ARDS. LPS was used to induce mice to establish ARDS model in vivo and to induce A549 cells to establish ARDS model in vitro. qRT-PCR was performed to determine the expressions of SNHG5, miR-205, and inflammatory cytokines. MTT assay was applied to detect cell viability. Dual-luciferase reporter (DLR) assay was performed to test the interactions among SNHG5, miR-205 and COMMD1. Western blot was used to detect the protein expression of COMMD1. Lung injury was evaluated by evaluating the score of lung injury, lung wet/dry weight ratio, and myeloperoxidase (MPO) activity. SNHG5 was downregulated, while miR-205 was upregulated in the serum of ARDS patients and lung tissues of LPS-induced mice. Upregulation of SNHG5 or down-regulation of miR-205 inhibited inflammation and promoted the viability of LPS-induced A549 cells. SNHG5 alleviated the lung injury of ARDS mice. MiR-205 was a target of SNHG5 and inversely correlated with SNHG5. COMMD1 was targeted by miR-205, and was positively regulated by SNHG5. MiR-205 mimics or sh-COMMD1 reversed the promoting effect of SNHG5 on cell viability and the suppressing effect of SNHG5 on inflammation in cellular model of ARDS. Meantime, miR-205 mimics reversed the relieving effect of SNHG5 on lung injury in mouse model of ARDS. SNHG5 acted as a sponge for miR-205 to ameliorate LPS-induced ARDS by regulating COMMD1.
先前的研究已经报道了长链非编码 RNA(lncRNA)在急性呼吸窘迫综合征(ARDS)中的重要作用。在这里,我们专注于 lncRNA SNHG5 在 ARDS 中的作用和调节机制。LPS 用于在体内诱导小鼠建立 ARDS 模型,并在体外诱导 A549 细胞建立 ARDS 模型。qRT-PCR 用于测定 SNHG5、miR-205 和炎症细胞因子的表达。MTT 测定法用于检测细胞活力。双荧光素酶报告(DLR)测定法用于测试 SNHG5、miR-205 和 COMMD1 之间的相互作用。Western blot 用于检测 COMMD1 蛋白表达。通过评估肺损伤评分、肺湿/干重比和髓过氧化物酶(MPO)活性来评估肺损伤。在 ARDS 患者的血清和 LPS 诱导的小鼠肺组织中,SNHG5 下调,而 miR-205 上调。上调 SNHG5 或下调 miR-205 抑制炎症并促进 LPS 诱导的 A549 细胞活力。SNHG5 减轻 ARDS 小鼠的肺损伤。miR-205 是 SNHG5 的靶标,与 SNHG5 呈负相关。COMMD1 是 miR-205 的靶标,受 SNHG5 正向调节。miR-205 模拟物或 sh-COMMD1 逆转了 SNHG5 对细胞活力的促进作用和 SNHG5 对细胞模型中炎症的抑制作用。同时,miR-205 模拟物逆转了 SNHG5 对 ARDS 小鼠模型中肺损伤的缓解作用。SNHG5 作为 miR-205 的海绵通过调节 COMMD1 来改善 LPS 诱导的 ARDS。